Medical devices containing rapamycin analogs

ABSTRACT

A medical device comprising a supporting structure capable of containing or supporting a pharmaceutically acceptable carrier or excipient, which carrier or excipient may contain one or more therapeutic agents or substances, with the carrier preferably including a coating on the surface thereof, and the coating containing the therapeutic substances, such as, for example, drugs. Supporting structures for the medical devices that are suitable for use in this invention include, but are not limited to, coronary stents, peripheral stents, catheters, arterio-venous grafts, by-pass grafts, and drug delivery balloons used in the vasculature. Drugs that are suitable for use in this invention include, but are not limited to Formula (I). This drug can be used in combination with another drug including those selected from anti-proliferative agents, anti-platelet agents, anti-inflammatory agents, anti-thrombotic agents, cytotoxic drugs, agents that inhibit cytokine or chemokine binding, cell de-differentiation inhibitors, anti-lipaedemic agents, matrix metalloproteinase inhibitors, cytostatic drugs, or combinations of these drugs.

[0001] This application is a continuation-in-part of U.S. Ser. No.09/950,307, filed Sep. 10, 2001, which is a continuation-in-part of U.S.Ser. No. 09/433,001, filed Nov. 2, 1999, which is a divisional of U.S.Ser. No. 09/159,945, filed Sep. 24, 1998, now U.S. Pat. No. 6,015,815,incorporated herein by reference.

TECHNICAL FIELD

[0002] The present invention relates to novel chemical compounds havingimmunomodulatory activity and synthetic intermediates useful for thepreparation of the novel compounds, and in particular to macrolideimmunomodulators. More particularly, the invention relates tosemisynthetic analogs of rapamycin, means for their preparation,pharmaceutical compositions containing such compounds, and methods oftreatment employing the same.

BACKGROUND OF THE INVENTION

[0003] The compound cyclosporine (cyclosporin A) has found wide usesince its introduction in the fields of organ transplantation andimmunomodulation, and has brought about a significant increase in thesuccess rate for transplantation procedures. Recently, several classesof macrocyclic compounds having potent immunomodulatory activity havebeen discovered. Okuhara et al., in European Patent Application No.184,162, published Jun. 11, 1986, disclose a number of macrocycliccompounds isolated from the genus Streptomyces, including theimmunosuppressant FK-506, a 23-membered macrocyclic lactone, which wasisolated from a strain of S. tsukubaensis.

[0004] Other related natural products, such as FR-900520 and FR-900523,which differ from FK-506 in their alkyl substituent at C-21, have beenisolated from S. hygroscopicus yakushimnaensis. Another analog,FR-900525, produced by S. tsukubaensis, differs from FK-506 in thereplacement of a pipecolic acid moiety with a proline group.Unsatisfactory side-effects associated with cyclosporine and FK-506 suchas nephrotoxicity, have led to a continued search for immunosuppressantcompounds having improved efficacy and safety, including animmunosuppressive agent which is effective topically, but ineffectivesystemically (U.S. Pat. No. 5,457,111).

[0005] Rapamycin is a macrocyclic triene antibiotic produced byStreptomyces hygroscopicus, which was found to have antifungal activity,particularly against Candida albicans, both in vitro and in vivo (C.Vezina et al., J. Antibiot. 1975, 28, 721; S. N. Sehgal et al., J.Antibiot 1975, 28, 727; H. A. Baker et al., J. Antibiot 1978, 31, 539;U.S. Pat. No. 3,929,992; and U.S. Pat. No. 3,993,749).

[0006] Rapamycin alone (U.S. Pat. No. 4,885,171) or in combination withpicibanil (U.S. Pat. No. 4,401,653) has been shown to have antitumoractivity. In 1977, rapamycin was also shown to be effective as animmunosuppressant in the experimental allergic encephalomyelitis model,a model for multiple sclerosis; in the adjuvant arthritis model, a modelfor rheumatoid arthritis; and was shown to effectively inhibit theformation of IgE-like antibodies (R. Martel et al., Can. J. Physiol.Pharmacol., 1977, 55, 48).

[0007] The immunosuppressive effects of rapamycin have also beendisclosed in FASEB, 1989, 3, 3411 as has its ability to prolong survivaltime of organ grafts in histoincompatible rodents (R. Morris, Med. Sci.Res., 1989, 17, 877). The ability of rapamycin to inhibit T-cellactivation was disclosed by M. Strauch (FASEB, 1989, 3, 3411). These andother biological effects of rapamycin are reviewed in TransplantationReviews, 1992, 6, 39-87.

[0008] Rapamycin has been shown to reduce neointimal proliferation inanimal models, and to reduce the rate of restenosis in humans. Evidencehas been published showing that rapamycin also exhibits ananti-inflammatory effect, a characteristic which supported its selectionas an agent for the treatment of rheumatoid arthritis. Because both cellproliferation and inflammation are thought to be causative factors inthe formation of restenotic lesions after balloon angioplasty and stentplacement, rapamycin and analogs thereof have been proposed for theprevention of restenosis.

[0009] Mono-ester and di-ester derivatives of rapamycin (esterificationat positions 31 and 42) have been shown to be useful as antifungalagents (U.S. Pat. No. 4,316,885) and as water soluble prodrugs ofrapamycin (U.S. Pat. No. 4,650,803).

[0010] Fermentation and purification of rapamycin and 30-demethoxyrapamycin have been described in the literature (C. Vezina et al. J.Antibiot. (Tokyo), 1975, 28 (10), 721; S. N. Sehgal et al., J. Antibiot(Tokyo), 1975, 28(10), 727; 1983, 36(4), 351; N. L. Pavia et al., J.Natural Products, 1991, 54(1), 167-177).

[0011] Numerous chemical modifications of rapamycin have been attempted.These include the preparation of mono- and di-ester derivatives ofrapamycin (WO 92/05179), 27-oximes of rapamycin (EPO 467606); 42-oxoanalog of rapamycin (U.S. Pat. No. 5,023,262); bicyclic rapamycins (U.S.Pat. No. 5,120,725); rapamycin dimers (U.S. Pat. No. 5,120,727); silylethers of rapamycin (U.S. Pat. No. 5,120,842); and arylsulfonates andsulfamates (U.S. Pat. No. 5,177,203). Rapamycin was recently synthesizedin its naturally occurring enantiomeric form (K. C. Nicolaou et al., J.Am. Chem. Soc., 1993, 115, 4419-4420; S. L. Schreiber, J. Am. Chem.Soc., 1993, 115, 7906-7907; S. J. Danishefsky, J. Am. Chem. Soc., 1993,115, 9345-9346.

[0012] It has been known that rapamycin, like FK-506, binds to FKBP-12(Siekierka, J. J.; Hung, S. H. Y.; Poe, M.; Lin, C. S.; Sigal, N. H.Nature, 1989, 341, 755-757; Harding, M. W.; Galat, A.; Uehling, D. E.;Schreiber, S. L. Nature 1989, 341, 758-760; Dumont, F. J.; Melino, M.R.; Staruch, M. J.; Koprak, S. L.; Fischer, P. A.; Sigal, N. H. J.Immunol. 1990, 144, 1418-1424; Bierer, B. E.; Schreiber, S. L.;Burakoff, S. J. Eur. J. Immunol. 1991, 21, 439-445; Fretz, H.; Albers,M. W.; Galat, A.; Standaert, R. F.; Lane, W. S.; Burakoff, S. J.;Bierer, B. E.; Schreiber, S. L. J. Am. Chem. Soc. 1991, 113, 1409-1411).Recently it has been discovered that the rapamycin/FKBP-12 complex bindsto yet another protein, which is distinct from calcineurin, the proteinthat the FK-506/FKBP-12 complex inhibits (Brown, E. J.; Albers, M. W.;Shin, T. B.; Ichikawa, K.; Keith, C. T.; Lane, W. S.; Schreiber, S. L.Nature 1994, 369, 756-758; Sabatini, D. M.; Erdjument-Bromage, H.; Lui,M.; Tempest, P.; Snyder, S. H. Cell, 1994, 78, 3543).

[0013] Percutaneous transluminal coronary angioplasty (PTCA) wasdeveloped by Andreas Gruntzig in the 1970's. The first canine coronarydilation was performed on Sep. 24, 1975; studies showing the use of PTCAwere presented at the annual meetings of the American Heart Associationthe following year. Shortly thereafter, the first human patient wasstudied in Zurich, Switzerland, followed by the first American humanpatients in San Francisco and New York. While this procedure changed thepractice of interventional cardiology with respect to treatment ofpatients with obstructive coronary artery disease, the procedure did notprovide long-term solutions. Patients received only temporary abatementof the chest pain associated with vascular occlusion; repeat procedureswere often necessary. It was determined that the existence of restenoticlesions severely limited the usefulness of the new procedure. In thelate 1980's, stents were introduced to maintain vessel patency afterangioplasty. Stenting is involved in 90% of angioplasty performed today.Before the introduction of stents, the rate of restenosis ranged from30% to 50% of the patients who were treated with balloon angioplasty.The recurrence rate after dilatation of in-stent restenosis may be ashigh as 70% in selected patient subsets, while the angiographicrestenosis rate in de novo stent placement is about 20%. Placement ofthe stent reduced the restenosis rate to 15% to 20%. This percentagelikely represents the best results obtainable with purely mechanicalstenting. The restenosis lesion is caused primarily by neointimalhyperplasia, which is distinctly different from atherosclerotic diseaseboth in time-course and in histopathologic appearance. Restenosis is ahealing process of damaged coronary arterial walls, with neointimaltissue impinging significantly on the vessel lumen. Vascularbrachytherapy appears to be efficacious against in-stent restenosislesions. Radiation, however, has limitations of practicality andexpense, and lingering questions about safety and durability.

[0014] Accordingly, it is desired to reduce the rate of restenosis by atleast 50% of its current level. It is for this reason that a majoreffort is underway by the interventional device community to fabricateand evaluate drug-eluting stents. Such devices could have manyadvantages if they were successful, principally since such systems wouldneed no auxiliary therapies, either in the form of peri-proceduraltechniques or chronic oral pharmacotherapy.

BRIEF DESCRIPTION OF THE DRAWINGS

[0015]FIG. 1 shows blood concentrations ±SEM (n=3) oftetrazole-containing rapamycin analogs dosed in monkey.

[0016]FIG. 2 is a side view in elevation showing a stent suitable foruse in this invention.

[0017]FIG. 3A is a cross-sectional view of a vessel segment in which wasplaced a stent coated with a polymer only.

[0018]FIG. 3B is a cross-sectional view of a vessel segment in which wasplaced a stent coated with a polymer plus drug.

SUMMARY OF THE INVENTION

[0019] In one aspect of the present invention are disclosed compoundsrepresented by the structural formula:

[0020] or a pharmaceutically acceptable salt or prodrug thereof.

[0021] Another object of the present invention is to provide syntheticprocesses for the preparation of such compounds from starting materialsobtained by fermentation, as well as chemical intermediates useful insuch synthetic processes.

[0022] A further object of the invention is to provide pharmaceuticalcompositions containing, as an active ingredient, at least one of theabove compounds.

[0023] Yet another object of the invention is to provide a method oftreating a variety of disease states, including restenosis,post-transplant tissue rejection, immune and autoimmune dysfunction,fungal growth, and cancer.

[0024] In another aspect this invention provides a medical devicecomprising a supporting structure having a coating on the surfacethereof, the coating containing a therapeutic substance, such as, forexample, a drug. Supporting structures for the medical devices that aresuitable for use in this invention include, but are not limited to,coronary stents, peripheral stents, catheters, arterio-venous grafts,by-pass grafts, and drug delivery balloons used in the vasculature.Drugs that are suitable for use in this invention include, but are notlimited to,

[0025] or a pharmaceutically acceptable salt or prodrug thereof, whichincludes

[0026] or a pharmaceutically acceptable salt or prodrug thereof,(hereinafter alternatively referred to as A-179578), and

[0027] or a pharmaceutically acceptable salt or prodrug thereof;

[0028] or a pharmaceutically acceptable salt or prodrug thereof,(hereinafter alternatively referred to as SDZ RAD or40-O-(2-hydroxyethyl)-rapamycin);

[0029] or a pharmaceutically acceptable salt or prodrug thereof,(hereinafter alternatively referred to as A-94507).

[0030] Coatings that are suitable for use in this invention include, butare not limited to, polymeric coatings that can comprise any polymericmaterial in which the therapeutic agent, i.e., the drug, issubstantially soluble. The coating can be hydrophilic, hydrophobic,biodegradable, or non-biodegradable. This medical device reducesrestenosis in vasculature. The direct coronary delivery of a drug suchas A-179578 is expected to reduce the rate of restenosis to a level ofabout 0% to 25%.

DETAILED DESCRIPTION OF THE INVENTION

[0031] Definition of Terms

[0032] The term “prodrug,” as used herein, refers to compounds which arerapidly transformed in vivo to the parent compound of the above formula,for example, by hydrolysis in blood. A thorough discussion is providedby T. Higuchi and V. Stella, “Pro-drugs as Novel Delivery systems,” Vol.14 of the A. C. S. Symposium Series, and in Edward B. Roche, ed.,“Bioreversible Carriers in Drug Design,” American PharmaceuticalAssociation and Pergamon Press, 1987, both of which are incorporatedherein by reference.

[0033] The term “pharmaceutically acceptable prodrugs”, as used herein,refers to those prodrugs of the compounds of the present invention whichare, within the scope of sound medical judgment, suitable for use incontact with the tissues of humans and lower mammals without unduetoxicity, irritation, and allergic response, are commensurate with areasonable benefit/risk ratio, and are effective for their intended use,as well as the zwitterionic forms, where possible, of the compounds ofthe invention. Particularly preferred pharmaceutically acceptableprodrugs of this invention are prodrug esters of the C-31 hydroxyl groupof compounds of this invention.

[0034] The term “prodrug esters,” as used herein, refers to any ofseveral ester-forming groups that are hydrolyzed under physiologicalconditions. Examples of prodrug ester groups include acetyl, ethanoyl,pivaloyl, pivaloyloxymethyl, acetoxymethyl, phthalidyl, methoxymethyl,indanyl, and the like, as well as ester groups derived from the couplingof naturally or unnaturally-occurring amino acids to the C-31 hydroxylgroup of compounds of this invention.

[0035] The term “supporting structure” means a framework that is capableof containing or supporting a pharmaceutically acceptable carrier orexcipient, which carrier or excipient may contain one or moretherapeutic agents or substances, e.g., one or more drugs and/or othercompounds. The supporting structure is typically formed of metal or apolymeric material. Suitable supporting structures formed of polymericmaterials, including biodegradable polymers, capable of containing thetherapeutic agents or substances include, without limitation, thosedisclosed in U.S. Pat. Nos. 6,413,272 and 5,527,337, which areincorporated herein by reference.

Embodiments

[0036] In one embodiment of the invention is a compound of formula

[0037] In another embodiment of the invention is a compound of formula

Preparation of Compounds of this Invention

[0038] The compounds and processes of the present invention will bebetter understood in connection with the following synthetic schemeswhich illustrate the methods by which the compounds of the invention maybe prepared.

[0039] The compounds of this invention may be prepared by a variety ofsynthetic routes. A representative procedure is shown in Scheme 1.

[0040] As shown in Scheme 1, conversion of the C-42 hydroxyl ofrapamycin to a trifluoromethanesulfonate or fluorosulfonate leavinggroup provided A. Displacement of the leaving group with tetrazole inthe presence of a hindered, non-nucleophilic base, such as 2,6-lutidine,or, preferably, diisopropylethyl amine provided epimers B and C, whichwere separated and purified by flash column chromatography.

Synthetic Methods

[0041] The foregoing may be better understood by reference to thefollowing examples which illustrate the methods by which the compoundsof the invention may be prepared and are not intended to limit the scopeof the invention as defined in the appended claims.

EXAMPLE 1 42-Epi-(tetrazolyl)-rapamycin (less polar isomer) Example 1A

[0042] A solution of rapamycin (100 mg, 0.11 mmol) in dichloromethane(0.6 mL) at −78° C. under a nitrogen atmosphere was treated sequentiallywith 2,6-lutidine (53 uL, 0.46 mmol, 4.3 eq.) andtrifluoromethanesulfonic anhydride (37 uL, 0.22 mmol), and stirredthereafter for 15 minutes, warmed to room temperature and eluted througha pad of silica gel (6 mL) with diethyl ether. Fractions containing thetriflate were pooled and concentrated to provide the designated compoundas an amber foam.

Example 1B 42-Epi-(tetrazolyl)-rapamycin (less polar isomer)

[0043] A solution of Example 1A in isopropyl acetate (0.3 mL) wastreated sequentially with diisopropylethylamine (87 mL, 0.5 mmol) and1H-tetrazole (35 mg, 0.5 mmol), and thereafter stirred for 18 hours.This mixture was partitioned between water (10 mL) and ether (10 mL).The organics were washed with brine (10 mL) and dried (Na₂SO₄).Concentration of the organics provided a sticky yellow solid which waspurified by chromatography on silica gel (3.5 g, 70-230 mesh) elutingwith hexane (10 mL), hexane:ether (4:1(10 mL), 3:1(10 mL), 2:1(10 mL),1:1(10 mL)), ether (30 mL), hexane:acetone (1:1(30 mL)). One of theisomers was collected in the ether fractions.

[0044] MS (ESI) m/e 966 (M)⁻;

EXAMPLE 2 42-Epi-(tetrazolyl)-rapamycin (more polar isomer) Example 2A42-Epi-(tetrazolyl)-rapamycin (more polar isomer)

[0045] Collection of the slower moving band from the chromatographycolumn using the hexane:acetone (1:1) mobile phase in Example 1Bprovided the designated compound.

[0046] MS (ESI) m/e 966 (M)⁻.

[0047] In Vitro Assay of Biological Activity

[0048] The immunosuppressant activity of the compounds of the presentinvention was compared to rapamycin and two rapamycin analogs:40-epi-N-[2′-pyridone]-rapamycin and 40-epi-N-[4′-pyridone]-rapamycin,both disclosed in U.S. Pat. No. 5,527,907. The activity was determinedusing the human mixed lymphocyte reaction (MLR) assay described by Kino,T. et al. in Transplantation Proceedings, XIX(5):36-39, Suppl. 6 (1987).The results of the assay demonstrate that the compounds of the inventionare effective immunomodulators at nanomolar concentrations, as shown inTable 1. TABLE 1 Human MLR Example IC₅₀ ± S.E.M.(nM) Rapamycin 0.91 ±0.36 2-pyridone 12.39 ± 5.3  4-pyridone 0.43 ± 0.20 Example 1 1.70 ±0.48 Example 2 0.66 ± 0.19

[0049] The pharmacokinetic behaviors of Example 1 and Example 2 werecharacterized following a single 2.5 mg/kg intravenous dose incynomolgus monkey (n=3 per group). Each compound was prepared as 2.5mg/mL solution in a 20% ethanol:30% propylene glycol:2% cremophor EL:48%dextrose 5% in water vehicle.

[0050] The 1 mL/kg intravenous dose was administered as a slow bolus(˜1-2 minutes) in a saphenous vein of the monkeys. Blood samples wereobtained from a femoral artery or vein of each animal prior to dosingand 0.1 (IV only), 0.25, 0.5, 1, 1.5, 2, 4, 6, 9, 12, 24, and 30 hoursafter dosing. The EDTA preserved samples were thoroughly mixed andextracted for subsequent analysis.

[0051] An aliquot of blood (1.0 mL) was hemolyzed with 20% methanol inwater (0.5 ml) containing an internal standard. The hemolyzed sampleswere extracted with a mixture of ethyl acetate and hexane (1:1 (v/v),6.0 mL). The organic layer was evaporated to dryness with a stream ofnitrogen at room temperature. Samples were reconstituted in methanol:water (1:1, 150 μL). The title compounds (50 μL injection) wereseparated from contaminants using reverse phase HPLC with UV detection.Samples were kept cool (4° C.) through the run. All samples from eachstudy were analyzed as a single batch on the HPLC.

[0052] Area under the curve (AUC) measurements of Example 1, Example 2and the internal standard were determined using the Sciex MacQuan™software. Calibration curves were derived from peak area ratio (parentdrug/internal standard) of the spiked blood standards using leastsquares linear regression of the ratio versus the theoreticalconcentration. The methods were linear for both compounds over the rangeof the standard curve (correlation >0.99) with an estimated quantitationlimit of 0.1 ng/mL. The maximum blood concentration (C_(MAX)) and thetime to reach the maximum blood concentration (T_(MAX)) were readdirectly from the observed blood concentration-time data. The bloodconcentration data were submitted to multi-exponential curve fittingusing CSTRIP to obtain estimates of pharmacokinetic parameters. Theestimated parameters were further defined using NONLIN84. The area underthe blood concentration-time curve from 0 to t hours (last measurableblood concentration time point) after dosing (AUC_(0-t)) was calculatedusing the linear trapeziodal rule for the blood-time profiles. Theresidual area extrapolated to infinity, determined as the final measuredblood concentration (C_(t)) divided by the terminal elimination rateconstant (β), and added to AUC_(0-t) to produce the total area under thecurve (AUC_(0-t)).

[0053] As shown in FIG. 1 and Table 2, both Example 1 and Example 2 hada surprisingly substantially shorter terminal elimination half-life(t_(1/2)) when compared to rapamycin. Thus, only the compounds of theinvention provide both sufficient efficacy (Table 1) and a shorterterminal half-life (Table 2). TABLE 2 AUC t_(1/2) Compound ng · hr/mL(hours) Rapamycin 6.87 16.7 2-pyridone 2.55 2.8 4-pyridone 5.59 13.3Example 1 2.35 5.0 Example 2 2.38 6.9

[0054] Methods of Treatment

[0055] The compounds of the invention, including but not limited tothose specified in the examples, possess immunomodulatory activity inmammals (especially humans). As immunosuppressants, the compounds of thepresent invention are useful for the treatment and prevention ofimmune-mediated diseases such as the resistance by transplantation oforgans or tissue such as heart, kidney, liver, medulla ossium, skin,cornea, lung, pancreas, intestinum tenue, limb, muscle, nerves,duodenum, small-bowel, pancreatic-islet-cell, and the like;graft-versus-host diseases brought about by medulla ossiumtransplantation; autoimmune diseases such as rheumatoid arthritis,systemic lupus erythematosus, Hashimoto's thyroiditis, multiplesclerosis, myasthenia gravis, type I diabetes, uveitis, allergicencephalomyelitis, glomerulonephritis, and the like. Further usesinclude the treatment and prophylaxis of inflammatory andhyperproliferative skin diseases and cutaneous manifestations ofimmunologically-mediated illnesses, such as psoriasis, atopicdermatitis, contact dermatitis and further eczematous dermatitises,seborrhoeis dermatitis, lichen planus, pemphigus, bullous pemphigoid,epidermolysis bullosa, urticaria, angioedemas, vasculitides, erythemas,cutaneous eosinophilias, lupus erythematosus, acne and alopecia areata;various eye diseases (autoimmune and otherwise) such askeratoconjunctivitis, vernal conjunctivitis, uveitis associated withBehcet's disease, keratitis, herpetic keratitis, conical cornea,dystrophia epithelialis corneae, corneal leukoma, and ocular pemphigus.In addition reversible obstructive airway disease, which includesconditions such as asthma (for example, bronchial asthma, allergicasthma, intrinsic asthma, extrinsic asthma and dust asthma),particularly chronic or inveterate asthma (for example, late asthma andairway hyper-responsiveness), bronchitis, allergic rhinitis, and thelike are targeted by compounds of this invention. Inflammation of mucosaand blood vessels such as gastric ulcers, vascular damage caused byischemic diseases and thrombosis. Moreover, hyperproliferative vasculardiseases such as intimal smooth muscle cell hyperplasia, restenosis andvascular occlusion, particularly following biologically- ormechanically-mediated vascular injury, could be treated or prevented bythe compounds of the invention.

[0056] The compounds or drugs described herein can be applied to stentsthat have been coated with a polymeric compound. Incorporation of thecompound or drug into the polymeric coating of the stent can be carriedout by dipping the polymer-coated stent into a solution containing thecompound or drug for a sufficient period of time (such as, for example,five minutes) and then drying the coated stent, preferably by means ofair drying for a sufficient period of time (such as, for example, 30minutes). The polymer-coated stent containing the compound or drug canthen be delivered to the coronary vessel by deployment from a ballooncatheter. In addition to stents, other devices that can be used tointroduce the drugs of this invention to the vasculature include, butare not limited to grafts, catheters, and balloons. In addition, othercompounds or drugs that can be used in lieu of the drugs of thisinvention include, but are not limited to, A-94507 and SDZ RAD).

[0057] The compounds described herein for use in polymer-coated stentscan be used in combination with other pharmacological agents. Thepharmacological agents that would, in combination with the compounds ofthis invention, be most effective in preventing restenosis can beclassified into the categories of anti-proliferative agents,anti-platelet agents, anti-inflammatory agents, anti-thrombotic agents,and thrombolytic agents. These classes can be further sub-divided. Forexample, anti-proliferative agents can be anti-mitotic. Anti-mitoticagents inhibit or affect cell division, whereby processes normallyinvolved in cell division do not take place. One sub-class ofanti-mitotic agents includes vinca alkaloids. Representative examples ofvinca alkaloids include, but are not limited to, vincristine,paclitaxel, etoposide, nocodazole, indirubin, and anthracyclinederivatives, such as, for example, daunorubicin, daunomycin, andplicamycin. Other sub-classes of anti-mitotic agents includeanti-mitotic alkylating agents, such as, for example, tauromustine,bofumustine, and fotemustine, and anti-mitotic metabolites, such as, forexample, methotrexate, fluorouracil, 5-bromodeoxyuridine, 6-azacytidine,and cytarabine. Anti-mitotic alkylating agents affect cell division bycovalently modifying DNA, RNA, or proteins, thereby inhibiting DNAreplication, RNA transcription, RNA translation, protein synthesis, orcombinations of the foregoing.

[0058] Anti-platelet agents are therapeutic entities that act by (1)inhibiting adhesion of platelets to a surface, typically a thrombogenicsurface, (2) inhibiting aggregation of platelets, (3) inhibitingactivation of platelets, or (4) combinations of the foregoing.Activation of platelets is a process whereby platelets are convertedfrom a quiescent, resting state to one in which platelets undergo anumber of morphologic changes induced by contact with a thrombogenicsurface. These changes include changes in the shape of the platelets,accompanied by the formation of pseudopods, binding to membranereceptors, and secretion of small molecules and proteins, such as, forexample, ADP and platelet factor 4. Anti-platelet agents that act asinhibitors of adhesion of platelets include, but are not limited to,eptifibatide, tirofiban, RGD (Arg-Gly-Asp)-based peptides that inhibitbinding to gpllbllla or αvβ3, antibodies that block binding to gpllalllbor αvβ3, anti-P-selectin antibodies, anti-E-selectin antibodies,peptides that block P-selectin or E-selectin binding to their respectiveligands, saratin, and anti-von Willebrand factor antibodies. Agents thatinhibit ADP-mediated platelet aggregation include, but are not limitedto, disagregin and cilostazol.

[0059] Anti-inflammatory agents can also be used. Examples of theseinclude, but are not limited to, prednisone, dexamethasone,hydrocortisone, estradiol, and non-steroidal anti-inflammatories, suchas, for example, acetaminophen, ibuprofen, naproxen, and sulindac. Otherexamples of these agents include those that inhibit binding of cytokinesor chemokines to the cognate receptors to inhibit pro-inflammatorysignals transduced by the cytokines or the chemokines.

[0060] Representative examples of these agents include, but are notlimited to, anti-IL1, anti-IL2, anti-IL3, anti-IL4, anti-IL8, anti-IL15,anti-GM-CSF, and anti-TNF antibodies.

[0061] Anti-thrombotic agents include chemical and biological entitiesthat can intervene at any stage in the coagulation pathway. Examples ofspecific entities include, but are not limited to, small molecules thatinhibit the activity of factor Xa. In addition, heparinoid-type agentsthat can inhibit both FXa and thrombin, either directly or indirectly,such as, for example, heparin, heparan sulfate, low molecular weightheparins, such as, for example, the compound having the trademarkClivarin®, and synthetic oligosaccharides, such as, for example, thecompound having the trademark Arixtra®. Also included are directthrombin inhibitors, such as, for example, melagatran, ximelagatran,argatroban, inogatran, and peptidomimetics of binding site of thePhe-Pro-Arg fibrinogen substrate for thrombin. Another class ofanti-thrombotic agents that can be delivered are factor VII/VIIainhibitors, such as, for example, anti-factor VII/VIIa antibodies,rNAPc2, and tissue factor pathway inhibitor (TFPI).

[0062] Thrombolytic agents, which may be defined as agents that helpdegrade thrombi (clots), can also be used as adjunctive agents, becausethe action of lysing a clot helps to disperse platelets trapped withinthe fibrin matrix of a thrombus. Representative examples of thrombolyticagents include, but are not limited to, urokinase or recombinanturokinase, pro-urokinase or recombinant pro-urokinase, tissueplasminogen activator or its recombinant form, and streptokinase.

[0063] Other drugs that can be used in combination with the compounds ofthis invention are cytotoxic drugs, such as, for example, apoptosisinducers, such as TGF, and topoisomerase inhibitors, such as,10-hydroxycamptothecin, irinotecan, and doxorubicin. Other classes ofdrugs that can be used in combination with the compounds of thisinvention are drugs that inhibit cell de-differentiation and cytostaticdrugs.

[0064] Other agents that can be used in combination with the compoundsof this invention include anti-lipaedemic agents, such as, for example,fenofibrate, matrix metalloproteinase inhibitors, such as, for example,batimistat, antagonists of the endothelin-A receptor, such as, forexample, darusentan, and antagonists of the α_(v)β₃ integrin receptor.

[0065] When used in the present invention, the coating can comprise anypolymeric material in which the therapeutic agent, i.e., the drug, issubstantially soluble. The purpose of the coating is to serve as acontrolled release vehicle for the therapeutic agent or as a reservoirfor a therapeutic agent to be delivered at the site of a lesion. Thecoating can be polymeric and can further be hydrophilic, hydrophobic,biodegradable, or non-biodegradable. The material for the polymericcoating can be selected from the group consisting of polycarboxylicacids, cellulosic polymers, gelatin, polyvinylpyrrolidone, maleicanhydride polymers, polyamides, polyvinyl alcohols, polyethylene oxides,glycosaminoglycans, polysaccharides, polyesters, polyurethanes,silicones, polyorthoesters, polyanhydrides, polycarbonates,polypropylenes, polylactic acids, polyglycolic acids, polycaprolactones,polyhydroxybutyrate valerates, polyacrylamides, polyethers, and mixturesand copolymers of the foregoing. Coatings prepared from polymericdispersions such as polyurethane dispersions (BAYHYDROL, etc.) andacrylic acid latex dispersions can also be used with the therapeuticagents of the present invention.

[0066] Biodegradable polymers that can be used in this invention includepolymers such as poly(L-lactic acid), poly(DL-lactic acid),polycaprolactone, poly(hydroxy butyrate), polyglycolide,poly(diaxanone), poly(hydroxy valerate), polyorthoester; copolymers suchas poly (lactide-co-glycolide), polyhydroxy (butyrate-co-valerate),polyglycolide-co-trimethylene carbonate; polyanhydrides;polyphosphoester; polyphosphoester-urethane; polyamino acids;polycyanoacrylates; biomolecules such as fibrin, fibrinogen, cellulose,starch, collagen and hyaluronic acid; and mixtures of the foregoing.Biostable materials that are suitable for use in this invention includepolymers such as polyurethane, silicones, polyesters, polyolefins,polyamides, polycaprolactam, polyimide, polyvinyl chloride, polyvinylmethyl ether, polyvinyl alcohol, acrylic polymers and copolymers,polyacrylonitrile, polystyrene copolymers of vinyl monomers with olefins(such as styrene acrylonitrile copolymers, ethylene methyl methacrylatecopolymers; ethylene vinyl acetate), polyethers, rayons, cellulosics(such as cellulose acetate, cellulose nitrate, cellulose propionate,etc.), parylene and derivatives thereof; and mixtures and copolymers ofthe foregoing.

[0067] Another polymer that can be used in this invention ispoly(MPC_(w):LAM_(x):HPMA_(y):TSMA_(z)) where w, x, y, and z representthe molar ratios of monomers used in the feed for preparing the polymerand MPC represents the unit 2-methacryoyloxyethylphosphorylcholine, LMArepresents the unit lauryl methacrylate, HPMA represents the unit2-hydroxypropyl methacrylate, and TSMA represents the unit3-trimethoxysilylpropyl methacrylate. The drug-impregnated stent can beused to maintain patency of a coronary artery previously occluded bythrombus and/or atherosclerotic plaque. The delivery of ananti-proliferative agent reduces the rate of in-stent restenosis.

[0068] Other treatable conditions include but are not limited toischemic bowel diseases, inflammatory bowel diseases, necrotizingenterocolitis, intestinal inflammations/allergies such as Coeliacdiseases, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn'sdisease and ulcerative colitis; nervous diseases such as multiplemyositis, Guillain-Barre syndrome, Meniere's disease, polyneuritis,multiple neuritis, mononeuritis and radiculopathy; endocrine diseasessuch as hyperthyroidism and Basedow's disease; hematic diseases such aspure red cell aplasia, aplastic anemia, hypoplastic anemia, idiopathicthrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis,pernicious anemia, megaloblastic anemia and anerythroplasia; bonediseases such as osteoporosis; respiratory diseases such as sarcoidosis,fibroid lung and idiopathic interstitial pneumonia; skin disease such asdermatomyositis, leukoderma vulgaris, ichthyosis vulgaris, photoallergicsensitivity and cutaneous T cell lymphoma; circulatory diseases such asarteriosclerosis, atherosclerosis, aortitis syndrome, polyarteritisnodosa and myocardosis; collagen diseases such as scleroderma, Wegener'sgranuloma and Sjogren's syndrome; adiposis; eosinophilic fasciitis;periodontal disease such as lesions of gingiva, periodontium, alveolarbone and substantia ossea dentis; nephrotic syndrome such asglomerulonephritis; male pattern alopecia or alopecia senilis bypreventing epilation or providing hair germination and/or promoting hairgeneration and hair growth; muscular dystrophy; Pyoderma and Sezary'ssyndrome; Addison's disease; active oxygen-mediated diseases, as forexample organ injury such as ischemia-reperfusion injury of organs (suchas heart, liver, kidney and digestive tract) which occurs uponpreservation, transplantation or ischemic disease (for example,thrombosis and cardiac infarction); intestinal diseases such asendotoxin-shock, pseudomembranous colitis and colitis caused by drug orradiation; renal diseases such as ischemic acute renal insufficiency andchronic renal insufficiency; pulmonary diseases such as toxinosis causedby lung-oxygen or drug (for example, paracort and bleomycins), lungcancer and pulmonary emphysema; ocular diseases such as cataracta,siderosis, retinitis, pigmentosa, senile macular degeneration, vitrealscarring and corneal alkali burn; dermatitis such as erythemamultiforme, linear IgA ballous dermatitis and cement dermatitis; andothers such as gingivitis, periodontitis, sepsis, pancreatitis, diseasescaused by environmental pollution (for example, air pollution), aging,carcinogenesis, metastasis of carcinoma and hypobaropathy; diseasescaused by histamine or leukotriene-C₄ release; Behcet's disease such asintestinal-, vasculo- or neuro-Behcet's disease, and also Behcet's whichaffects the oral cavity, skin, eye, vulva, articulation, epididymis,lung, kidney and so on. Furthermore, the compounds of the invention areuseful for the treatment and prevention of hepatic disease such asimmunogenic diseases (for example, chronic autoimmune liver diseasessuch as autoimmune hepatitis, primary biliary cirrhosis and sclerosingcholangitis), partial liver resection, acute liver necrosis (e.g.necrosis caused by toxin, viral hepatitis, shock or anoxia), B-virushepatitis, non-A/non-B hepatitis, cirrhosis (such as alcoholiccirrhosis) and hepatic failure such as fulminant hepatic failure,late-onset hepatic failure and “acute-on-chronic” liver failure (acuteliver failure on chronic liver diseases), and moreover are useful forvarious diseases because of their useful activity such as augmention ofchemotherapeutic effect, cytomegalovirus infection, particularly HCMVinfection, anti-inflammatory activity, sclerosing and fibrotic diseasessuch as nephrosis, scleroderma, pulmonary fibrosis, arteriosclerosis,congestive heart failure, ventricular hypertrophy, post-surgicaladhesions and scarring, stroke, myocardial infarction and injuryassociated with ischemia and reperfusion, and the like.

[0069] Additionally, compounds of the invention possess FK-506antagonistic properties. The compounds of the present invention may thusbe used in the treatment of immunodepression or a disorder involvingimmunodepression.

[0070] Examples of disorders involving immunodepression include AIDS,cancer, fungal infections, senile dementia, trauma (including woundhealing, surgery and shock) chronic bacterial infection, and certaincentral nervous system disorders. The immunodepression to be treated maybe caused by an overdose of an immunosuppressive macrocyclic compound,for example derivatives of12-(2-cyclohexyl-1-methylvinyl)-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene such as FK-506 or rapamycin. The overdosing of suchmedicaments by patients is quite common upon their realizing that theyhave forgotten to take their medication at the prescribed time and canlead to serious side effects.

[0071] The ability of the compounds of the invention to treatproliferative diseases can be demonstrated according to the methodsdescribed in Bunchman ET and CA Brookshire, Transplantation Proceed. 23967-968 (1991); Yamagishi, et al., Biochem. Biophys. Res. Comm. 191840-846 (1993); and Shichiri, et al., J. Clin. Invest. 87 1867-1871(1991). Proliferative diseases include smooth muscle proliferation,systemic sclerosis, cirrhosis of the liver, adult respiratory distresssyndrome, idiopathic cardiomyopathy, lupus erythematosus, diabeticretinopathy or other retinopathies, psoriasis, scleroderma, prostatichyperplasia, cardiac hyperplasia, restenosis following arterial injuryor other pathologic stenosis of blood vessels. In addition, thesecompounds antagonize cellular responses to several growth factors, andtherefore possess antiangiogenic properties, making them useful agentsto control or reverse the growth of certain tumors, as well as fibroticdiseases of the lung, liver, and kidney.

[0072] Aqueous liquid compositions of the present invention areparticularly useful for the treatment and prevention of various diseasesof the eye such as autoimmune diseases (including, for example, conicalcornea, keratitis, dysophia epithelialis corneae, leukoma, Mooren'sulcer, sclevitis and Graves' ophthalmopathy) and rejection of cornealtransplantation.

[0073] When used in the above or other treatments, a therapeuticallyeffective amount of one of the compounds of the present invention may beemployed in pure form or, where such forms exist, in pharmaceuticallyacceptable salt, ester or prodrug form. Alternatively, the compound maybe administered as a pharmaceutical composition containing the compoundof interest in combination with one or more pharmaceutically acceptableexcipients. The phrase “therapeutically effective amount” of thecompound of the invention means a sufficient amount of the compound totreat disorders, at a reasonable benefit/risk ratio applicable to anymedical treatment. It will be understood, however, that the total dailyusage of the compounds and compositions of the present invention will bedecided by the attending physician within the scope of sound medicaljudgment. The specific therapeutically effective dose level for anyparticular patient will depend upon a variety of factors including thedisorder being treated and the severity of the disorder; activity of thespecific compound employed; the specific composition employed; the age,body weight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed; andlike factors well known in the medical arts. For example, it is wellwithin the skill of the art to start doses of the compound at levelslower than required to achieve the desired therapeutic effect and togradually increase the dosage until the desired effect is achieved.

[0074] The total daily dose of the compounds of this inventionadministered to a human or lower animal may range from about 0.01 toabout 10 mg/kg/day. For purposes of oral administration, more preferabledoses may be in the range of from about 0.001 to about 3 mg/kg/day. Forthe purposes of local delivery from a stent, the daily dose that apatient will receive depends on the length of the stent. For example, a15 mm coronary stent may contain a drug in an amount ranging from about1 to about 120 micrograms and may deliver that drug over a time periodranging from several hours to several weeks. If desired, the effectivedaily dose may be divided into multiple doses for purposes ofadministration; consequently, single dose compositions may contain suchamounts or submultiples thereof to make up the daily dose. Topicaladministration may involve doses ranging from 0.001 to 3% mg/kg/day,depending on the site of application.

[0075] Pharmaceutical Compositions

[0076] The pharmaceutical compositions of the present invention comprisea compound of the invention and a pharmaceutically acceptable carrier orexcipient, which may be administered orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, drops or transdermal patch), bucally, as an oral ornasal spray, or locally, as in a stent placed within the vasculature.The phrase “pharmaceutically acceptable carrier” means a non-toxicsolid, semi-solid or liquid filler, diluent, encapsulating material orformulation auxiliary of any type. The term “parenteral,” as usedherein, refers to modes of administration which include intravenous,intraarterial, intramuscular, intraperitoneal, intrasternal,subcutaneous and intraarticular injection, infusion, and placement, suchas, for example, in vasculature.

[0077] Pharmaceutical compositions of this invention for parenteralinjection comprise pharmaceutically acceptable sterile aqueous ornonaqueous solutions, dispersions, suspensions or emulsions as well assterile powders for reconstitution into sterile injectable solutions ordispersions just prior to use. Examples of suitable aqueous andnonaqueous carriers, diluents, solvents or vehicles include water,ethanol, polyols (such as glycerol, propylene glycol, polyethyleneglycol, and the like), carboxymethylcellulose and suitable mixturesthereof, vegetable oils (such as olive oil), and injectable organicesters such as ethyl oleate. Proper fluidity can be maintained, forexample, by the use of coating materials such as lecithin, by themaintenance of the required particle size in the case of dispersions,and by the use of surfactants.

[0078] These compositions may also contain adjuvants such aspreservatives, wetting agents, emulsifying agents, and dispersingagents. Prevention of the action of microorganisms may be ensured by theinclusion of various antibacterial and antifungal agents, for example,paraben, chlorobutanol, phenol sorbic acid, and the like. It may also bedesirable to include isotonic agents such as sugars, sodium chloride,and the like. Prolonged absorption of the injectable pharmaceutical formmay be brought about by the inclusion of agents that delay absorptionsuch as aluminum monostearate and gelatin.

[0079] In some cases, in order to prolong the effect of the drug, it isdesirable to slow the absorption of the drug from subcutaneous orintramuscular injection. This may be accomplished by the use of a liquidsuspension of crystalline or amorphous material with poor watersolubility. The rate of absorption of the drug then depends upon itsrate of dissolution which, in turn, may depend upon crystal size andcrystalline form. Alternatively, delayed absorption of a parenterallyadministered drug form is accomplished by dissolving or suspending thedrug in an oil vehicle.

[0080] Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissues.

[0081] The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium just prior to use.

[0082] Solid dosage forms for oral administration include capsules,tablets, pills, powders, and granules. In such solid dosage forms, theactive compound is mixed with at least one inert, pharmaceuticallyacceptable excipient or carrier such as sodium citrate or dicalciumphosphate and/or a) fillers or extenders such as starches, lactose,sucrose, glucose, mannitol, and silicic acid, b) binders such as, forexample, carboxymethylcellulose, alginates, gelatin,polyvinylpyrrolidone, sucrose, and acacia, c) humectants such asglycerol, d) disintegrating agents such as agar-agar, calcium carbonate,potato or tapioca starch, alginic acid, certain silicates, and sodiumcarbonate, e) solution retarding agents such as paraffin, f) absorptionaccelerators such as quaternary ammonium compounds, g) wetting agentssuch as, for example, cetyl alcohol and glycerol monostearate, h)absorbents such as kaolin and bentonite clay, and i) lubricants such astalc, calcium stearate, magnesium stearate, solid polyethylene glycols,sodium lauryl sulfate, and mixtures thereof. In the case of capsules,tablets and pills, the dosage form may also comprise buffering agents.

[0083] Solid compositions of a similar type may also be employed asfillers in soft, semi-solid and hard-filled gelatin capsules orliquid-filled capsules using such excipients as lactose or milk sugar aswell as high molecular weight polyethylene glycols and the like.

[0084] The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Those embedding compositionscontaining a drug can be placed on medical devices, such as stents,grafts, catheters, and balloons.

[0085] The active compounds can also be in micro-encapsulated form, ifappropriate, with one or more of the above-mentioned excipients.

[0086] Liquid dosage forms for oral administration includepharmaceutically acceptable emulsions, solutions, suspensions, syrupsand elixirs. In addition to the active compounds, the liquid dosageforms may contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethyl formamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof.

[0087] Besides inert diluents, the oral compositions can also includeadjuvants such as wetting agents, emulsifying and suspending agents,sweetening, flavoring, and perfuming agents.

[0088] Suspensions, in addition to the active compounds, may containsuspending agents as, for example, ethoxylated isostearyl alcohols,polyoxyethylene sorbitol an sorbitan esters, microcrystalline cellulose,aluminum metahydroxide, bentonite, aga agar, and tragacanth, andmixtures thereof.

[0089] Topical administration includes administration to the skin ormucosa, includin surfaces of the lung and eye. Compositions for topicaladministration, including those for inhalation, may be prepared as a drypowder which may be pressurized or non-pressurized. In non-pressurizedpowder compositions, the active ingredient in finely divided form may beused in admixture with a larger-sized pharmaceutically acceptable inertcarrier comprising particles having a size, for example, of up to 100micrometers in diameter. Suitable inert carriers include sugars such aslactose. Desirably, at least 95% by weight of the particles of theactive ingredient have an effective particle size in the range of 0.01to 10 micrometers. Compositions for topical use on the skin also includeointments, creams, lotions, and gels.

[0090] Alternatively, the composition may be pressurized and contain acompressed gas, such as nitrogen or a liquefied gas propellant. Theliquefied propellant mediurr and indeed the total composition ispreferably such that the active ingredient does not dissolve therein toany substantial extent. The pressurized composition may als contain asurface active agent. The surface active agent may be a liquid or solidnon-ionic surface active agent or may be a solid anionic surface activeagent. It is preferred to use the solid anionic surface active agent inthe form of a sodium salt.

[0091] A further form of topical administration is to the eye, as forthe treatment of immune-mediated conditions of the eye such asautoimmune diseases, allergic or inflammatory conditions, and cornealtransplants. The compound of the invention is delivered in apharmaceutically acceptable ophthalmic vehicle, such that the compoundis maintained in contact with the ocular surface for a sufficient timeperiod to allow the compound to penetrate the corneal and internalregions of the eye, as for example the anterior chamber, posteriorchamber, vitreous body, aqueous humor, vitreous humor, cornea,iris/cilary, lens, choroid/retina and sclera. The pharmaceuticallyacceptable ophthalmic vehicle may, for example, be an ointment,vegetable oil or an encapsulating material.

[0092] Compositions for rectal or vaginal administration are preferablysuppositories or retention enemas which can be prepared by mixing thecompounds of this invention with suitable non-irritating excipients orcarriers such as cocoa butter, polyethylene glycol or a suppository waxwhich are solid at room temperature but liquid at body temperature andtherefore melt in the rectum or vaginal cavity and release the activecompound.

[0093] Compounds of the present invention can also be administered inthe form of liposomes. As is known in the art, liposomes are generallyderived from phospholipids or other lipid substances. Liposomes areformed by mono- or multi-lamellar hydrated liquid crystals that aredispersed in an aqueous medium. Any non-toxic, physiologicallyacceptable and metabolizable lipid capable of forming liposomes can beused. The present compositions in liposome form can contain, in additionto a compound of the present invention, stabilizers, preservatives,excipients, and the like. The preferred lipids are the phospholipids andthe phosphatidyl cholines (lecithins), both natural and synthetic.Methods to form liposomes are known in the art. See, for example,Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, NewYork, N.Y. (1976), p. 33 et seq.

[0094] Compounds of the present invention may also be coadministeredwith one or more immunosuppressant agents. The immunosuppressant agentswithin the scope of this invention include, but are not limited to,IMURAN® azathioprine sodium, brequinar sodium, SPANIIDIN® gusperimustrihydrochloride (also known as deoxyspergualin), mizoribine (also knownas bredinin), CELLCEPT® mycophenolate mofetil, NEORAL® Cylosporin A(also marketed as different formulation of Cyclosporin A under thetrademark SANDIMMUNE®), PROGRAF® tacrolimus (also known as FK-506),sirolimus and RAPAMUNE®, leflunomide (also known as HWA-486),glucocorticoids, such as prednisolone and its derivatives, antibodytherapies such as orthoclone (OKT3) and Zenapax®, and antithymyocyteglobulins, such as thymoglobulins.

EXAMPLE 3

[0095] The purpose of this example was to determine the effects of arapamycin analog on neointimal formation in porcine coronary arteriescontaining stents. This example illustrates that the rapamycin analogA-179578, when compounded and delivered from the BiocompatiblesBiodiviYsio PC Coronary stent favorably affects neointimal hyperplasiaand lumen size in porcine coronary arteries. This finding suggests thatsuch a combination may be of substantial clinical benefit if properlyapplied in humans by limiting neointimal hyperplasia.

[0096] The agent A-179578 is a rapamycin analog. The study set forth inthis example was designed to assess the ability of the rapamycin analogA-179578 to reduce neointimal hyperplasia in a porcine coronary stentmodel. Efficacy of A-179578 in this model would suggest its clinicalpotential for the limitation and treatment of coronary restenosis instents following percutaneous revascularization. The domestic swine wasused because this model appears to yield results comparable to otherinvestigations seeking to limit neointimal hyperplasia in humansubjects.

[0097] The example tested A-179578 eluted from coronary stents placed injuvenile farm pigs, and compared these results with control stents. Thecontrol stents had polymer alone covering its struts. This is important,for the polymer itself must not stimulate neointimal hyperplasia to asubstantial degree. As the eluted drug disappears, an inflammatoryresponse to the polymer could conceivably result in a late “catch-upphenomenon” where the restenosis process is not stopped, but insteadslowed. This phenomenon would result in restenosis at late dates inhuman subjects.

[0098] Stents were implanted in two blood vessels in each pig. Pigs usedin this model were generally 24 months old and weighed 3040 Kg. Twocoronary stents were thus implanted in each pig by visually assessing aAnormal@ stent:artery ratio of 1.1-1.2.

[0099] Beginning on the day of the procedure, pigs were given oralaspirin (325 mg daily) and continued for the remainder of their course.General anesthesia was achieved by means of intramuscular injectionfollowed by intravenous ketamine (30 mg/kg) and xylazine (3 mg/kg).Additional medication at the time of induction included atropine (1 mg)and flocillin (1 g) administered intramuscularly. During the stentingprocedure, an intraarterial bolus of 10,000 units of heparin wasadministered.

[0100] Arterial access was obtained by cutdown on the right externalcarotid and placement of an 8F sheath. After the procedure, the animalswere maintained on a normal diet without cholesterol or other specialsupplementation.

[0101] The BiodivYsio stent was used with nominal vessel target size of3.0 mm. See FIG. 2. Two coronary arteries per pig were assigned atrandom to deployment of the stents. The stent was either a drug elutingstent (polymer plus drug stent) or a stent coated with a polymer only(polymer only stent). The stents were delivered by means of standardguide catheters and wires. The stent balloons were inflated toappropriate sizes for less than 30 seconds.

[0102] Each pig had one polymer only stent and one polymer plus drugstent placed in separate coronary arteries, so that each pig would haveone stent for drug and one for control.

[0103] A sample size of 20 pigs total was chosen to detect a projecteddifference in neointimal thickness of 0.2 mm with a standard deviationof 0.15 mm, at a power of 0.95 and beta 0.02.

[0104] Animals were euthanized at 28 days for histopathologicexamination and quantification. Following removal of the heart from theperfusion pump system, the left atrial appendage was removed for accessto the proximal coronary arteries. Coronary arterial segments withinjuries were dissected free of the epicardium. Segments containinglesions was isolated, thereby allowing sufficient tissue to containuninvolved blood vessel at either end. The foregoing segments, eachroughly 2.5 cm in length, were embedded and processed by means ofstandard plastic embedding techniques. The tissues were subsequentlyprocessed and stained with hematoxylin-eosin and elastic-van Giesontechniques.

[0105] Low and high power light microscopy were used to make lengthmeasurements in the plane of microscopic view by means of a calibratedreticle and a digital microscopy system connected to a computeremploying calibrated analysis software.

[0106] The severity of vessel injury and the neointimal response weremeasured by calibrated digital microscopy. The importance of theintegrity of the internal elastic lamina is well-known to those skilledin the art. A histopathologic injury score in stented blood vessels hasbeen validated as being closely related to neointimal thickness. Thisscore is related to depth of injury and is as follows: Score Descriptionof Injury 0 Internal elastic lamina intact; endothelium typicallydenuded, media compressed but not lacerated. 1 Internal elastic laminalacerated; media typically compressed but not lacerated. 2 Internalelastic lacerated; media visibly lacerated; external elastic laminaintact but compressed. 3 External elastic lamina lacerated; typicallylarge lacerations of media extending through the external elasticlamina; coil wires sometimes residing in adventitia.

[0107] This quantitative measurement of injury was assessed for allstent wires of each stent section. The calibrated digital image was alsoused to measure at each stent wire site the neointimal thickness. Lumenarea, area contained with the internal elastic lamina, and area withinthe external elastic lamina were also measured.

[0108] At each stent wire site for a given section, the neointimalthickness was averaged to obtain a mean injury score for each section.The measurement of neointimal thickness was made to the abluminal sideof the stent wire, because the neointimal in all cases includes thisthickness.

[0109] The mid-stent segment was used for measurement, analysis, andcomparison. Data were also recorded (and included in the data section ofthis report) for proximal and distal segments.

[0110] The data analysis methods for this study did not need to takeinto account variable arterial injury across treatment/control groups,because mild to moderate injury is sensitive enough to detect treatmentdifferences. Paired t-testing was performed to compare variables acrossthe polymer only stents (control group) and polymer plus drug stents(treatment group). No animal died in this study before scheduledtimepoints.

[0111] Table 3 shows the pigs and arteries used. In Table 3, LCX meansthe circumflex branch of the left coronary artery, LAD means the leftanterior descending coronary artery, and RCA means the right coronaryartery. TABLE 3 Pigs and Vessels Used 1 2000-G-693 RCA - Control2000-G-693 LCX - Test 2 2000-G-698 RCA - Test 2000-G-698 LAD - Control 32000-G-702 RCA - Test 2000-G-702 LAD - Control 4 2000-G-709 RCA -Control 2000-G-709 LAD - Test 5 2000-G-306 RCA - Control 2000-G-306LAD - Test 2000-G-306 *LCX - Test 6 2000-G-672 RCA - Test 2000-G-672LAD - Control 7 2000-G-712 RCA - Control 2000-G-712 LCX - Test 82000-G-735 RCA - Control 2000-G-735 LAD - Test 9 2000-G-736 RCA -Control 2000-G-736 LCX - Test 10 2000-G-740 RCA - Test 2000-G-740 LAD -Control 11 2000-G-742 LAD - Test 2000-G-742 OM (LCX) - Control 122000-G-744 RCA - Test 2000-G-744 LAD - Control 13 2000-G-748 RCA - Test2000-G-748 LAD - Control 14 2000-G-749 RCA - Control 2000-G-749 LCX -Test 15 2000-G-753 RCA - Control 2000-G-753 LAD - Test 16 2000-G-754RCA - Test 2000-G-754 LCX - Control 17 2000-G-755 RCA - Control2000-G-755 LAD - Test 18 2000-G-756 RCA - Test 2000-G-756 LAD - Control19 2000-G-757 LAD - Control 2000-G-757 LCX - Test 20 2000-G-760 LAD -Test 2000-G-760 LCX - Control

[0112] Table 4 shows the summary results for all data for mean injuryand neointimal thickness for each stent, including proximal, mid, anddistal segments. Table 4 also shows lumen size, percent stenosis, andartery size as measured by the internal elastic laminae (IEL) andexternal elastic laminae (EEL). TABLE 4 Summary: All Measures (Distal,Mid, Proximal) mean % Neointimal ID prox ref dist ref lumen IEL EELinjury stenosis area NIT Control Distal Mean 4.46 3.96 4.88 7.66 9.000.22 36.10 2.79 0.41 SD 1.20 1.16 1.30 1.15 1.10 0.26 15.41 1.29 0.17Control Mid Mean 4.46 3.96 4.94 7.71 9.08 0.08 36.23 2.77 0.38 SD 1.201.16 1.44 1.07 1.15 0.14 14.93 1.20 0.16 Control Proximal Mean 4.46 3.965.11 7.89 9.30 0.15 35.35 2.78 0.38 SD 1.20 1.16 1.38 1.33 1.42 0.2211.94 1.04 0.12 Test Distal Mean 4.26 3.41 6.04 7.70 9.01 0.26 22.351.66 0.25 SD 1.26 0.96 1.55 1.49 1.47 0.43 8.58 0.58 0.06 Test Mid Mean4.26 3.41 6.35 7.75 8.98 0.04 18.71 1.41 0.22 SD 1.26 0.96 1.29 1.181.31 0.07 5.68 0.33 0.05 Test Proximal Mean 2.56 2.15 3.31 4.06 4.660.19 16.79 1.29 0.18 SD 1.66 1.37 2.39 3.48 4.15 0.13 9.97 0.80 0.12

[0113] There was no statistically significant difference for neointimalarea or thickness across proximal, mid, or distal segments within thetest group (polymer plus drug stents) or control groups (polymer onlystents). This observation is quite consistent with prior studies, andthus allows use of only the mid segment for statistical comparison oftest devices (polymer plus drug stents) vs. control devices (polymeronly stents).

[0114] Table 5 shows the statistical t-test comparisons across testgroups and control groups. There was a statistically significantdifference in neointimal thickness, neointimal area, lumen size, andpercent lumen stenosis, the drug eluting stent being clearly favored.Conversely, there were no statistically significant differences betweenthe test group (polymer plus drug stents) and the control group (polymeronly stents) for mean injury score, external elastic laminae, orinternal elastic laminae areas. TABLE 5 Statistical Comparison of Testvs. Control Parameters: Mid-Section Data t-test Statistics ParameterDifference t-test DF Std Error Lower 95% Upper 95% p Lumen −1.17 −2.2838 0.52 −2.21 −0.13 0.029 IEL 0.03 0.088 38 0.36 −0.71 0.78 0.93 EEL 0.20.499 38 0.39 −0.599 0.99 0.62 NI Thickness 0.18 5.153 38 0.034 0.1060.244 <.0001 NI Area 1.21 3.62 38 0.33 0.53 1.88 0.0008 Mean Injury0.038 1.137 38 0.033 −0.02 0.106 0.26 % Stenosis 14.54 2.97 38 4.9 4.6124.47 0.005

[0115] The reference arteries proximal and distal to the stentedsegments were observed, and quantitated. These vessels appeared normalin all cases, uninjured in both the control group (polymer only stents)and the test group (polymer plus drug stents). See FIGS. 3A and 3B. Thedata below show there were no statistically significant differences insize between the stents in the control group and the stents in the testgroup. Proximal Reference Distal Reference Diameter (mm) Diameter (mm)Control 4.46 ± 1.20 3.96 ± 1.16 (mean ± SD) Test 4.26 ± 1.26 3.41 ± 0.96(mean + SD)

[0116] The data suggest that statistically significant differencesexist, and these differences favor the stent that elutes A-179578. Thestent of this invention results in lower neointimal area, lowerneointimal thickness, and greater lumen area. There were no significantdifferences within the test group (polymer plus drug stents) and thecontrol group (polymer only stents) for neointimal or injury parameters.There were no significant differences in artery sizes (including thestent) for the control group compared to the test group. These latterfindings suggest no significant difference in the arterial remodelingcharacteristics of the polymeric coating containing the drug.

[0117] At most, mild inflammation was found on both the polymer plusdrug stent and the polymer only stent. This finding suggests that thepolymer exhibits satisfactory biocompatibility, even without drugloading. Other studies show that when drug has completely gone from thepolymer, the polymer itself creates enough inflammation to causeneointima. This phenomenon may be responsible for the late Acatch-up@phenomenon of clinical late restenosis. Because the polymer in thisexample did not cause inflammation in the coronary arteries, lateproblems related to the polymer after the drug is exhausted areunlikely.

[0118] In conclusion, a stent containing the compound A-179578 with apolymer showed a reduction in neointimal hyperplasia in the porcinemodel when placed in a coronary artery.

EXAMPLE 4

[0119] The purpose of this example is to determine the rate of releaseof the A-179578 drug from 316L Electropolished Stainless Steel Couponscoated with a biocompatible polymer containing phosphorylcholine sidegroups.

[0120] Rubber septa from lids from HPLC vials were removed from thevials and placed into glass vials so that the “Teflon” side faced up.These septa served as supports for the test samples. The test sampleswere 316L stainless steel coupons that had been previously coated with abiocompatible polymer containing phosphorylcholine side groups (PCpolymer). Coronary stents are commonly made of 316L stainless steel andcan be coated with the PC polymer to provide a depot site for loadingdrugs. The coated coupons, which serve to simulate stents, were placedonto the septa. By using a glass Hamilton Syringe, a solution ofA-179578 and ethanol (10 μl) was applied to the surface of each coupon.The solution contained A-179578 (30.6 mg) dissolved in 100% ethanol (3.0ml). The syringe was cleaned with ethanol between each application. Thecap to the glass vial was placed on the vial loosely, thereby assuringproper ventilation. The coupon was allowed to dry for a minimum of 1.5hours. Twelve (12) coupons were loaded in this way—six being used todetermine the average amount of drug loaded onto the device and sixbeing used to measure the time needed to release the drug from thedevices.

[0121] To determine the total amount of A-179578 loaded onto a coupon, acoupon was removed from the vial and placed into 50/50 acetonitrile/0.01M phosphate buffer (pH 6.0, 5.0 ml). The coupon was placed onto a 5210Branson sonicator for one hour. The coupon was then removed from thesolution, and the solution was assayed by HPLC.

[0122] The time release studies were performed by immersing and removingthe individual coupons from fresh aliquots (10.0 ml) of 0.01 M phosphatebuffer at a pH of 6.0 at each of the following time intervals—5, 15, 30and 60 minutes. For the remaining time points of 120, 180, 240, 300, 360minutes, volumes of 5.0 ml of buffer were used. To facilitate mixingduring the drug release phase, the samples were placed onto an Eberbachshaker set at low speed. All solution aliquots were assayed by HPLCafter the testing of the last sample was completed.

[0123] The HPLC analysis was performed with a Hewlett Packard series1100 instrument having the following settings: Injection Volume =  100μl Acquisition Time =   40 minutes Flow Rate =  1.0 ml/min ColumnTemperature = 40° C. Wavelength =  278 nm Mobile Phase = 65%Acetonitrile/35% H₂O Column = YMC ODS-A S5 μm, 4.6 × 250 mm Part No.A12052546WT

[0124] The results from the above experiment showed the followingrelease data: TABLE 6 Time Percent Standard (min.) Release Deviation0.00 0.00 0.00 5.00 1.87 1.12 15.00 2.97 1.47 30.00 3.24 1.28 60.00 3.291.29 120.00 3.92 1.28 180.00 4.36 1.33 240.00 4.37 1.35 300.00 6.34 2.07360.00 7.88 1.01

EXAMPLE 5

[0125] The purpose of this example was to determine the loading andrelease of A-179578 from 15 mm BiodivYsio drug delivery stents.

[0126] To load the stents with drug, a solution of A-179578 in ethanolat a concentration of 50 mg/ml was prepared and dispensed into twelvevials. Twelve individual polymer-coated stents were placed on fixturesdesigned to hold the stent in a vertical position and the stents wereimmersed vertically in the drug solution for five minutes. The stentsand fixtures were removed from the vials and excess drug solution wasblotted away by contacting the stents with an absorbent material. Thestents were then allowed to dry in air for 30 minutes in an invertedvertical position.

[0127] The stents were removed from the fixtures, and each stent wasplaced into 50/50 acetonitrile/phosphate buffer (pH 5.1, 2.0 ml) andsonicated for one hour. The stents were removed from the solution andsolutions were assayed for concentration of drug, which allowedcalculation of the amount of drug originally on the stents. This methodwas independently shown to remove at least 95% of the drug from thestent coating. On average, the stents contained 60 micrograms of drug±20 micrograms.

[0128] The drug-loaded stents were placed on the fixtures and placedinto 0.01 M phosphate buffer (pH=6.0, 1.9 ml) in individual vials. Thesesamples were placed onto a Eberbach shaker set at low speed to provideback-and-forth agitation. To avoid approaching drug saturation in thebuffer, the stents were transferred periodically to fresh buffer vialsat the following points: 15, 30, 45, 60, 120, 135, 150, 165, 180, 240,390 minutes. The dissolution buffer vials were assayed by HPLC for thedrug concentration at the end of the drug release period studied. Thedata, represented as % cumulative release of the drug as a function oftime, is shown in tabular form below: TABLE 7 Time (min) % CumulativeRelease of Drug  15 0.3  30 1.1  45 2.1  60 3.2 120 4.3 135 5.9 150 6.3165 6.8 180 7.4 240 10.8 390 13.2

EXAMPLE 6

[0129] The purpose of this example was to evaluate the safety andefficacy of different drug dosages on neointima formation. Drug wasdelivered from the BiodivYsio OC stent (15 mm) coated with A-179578.In-stent neointima formation was measured at four time intervals—3 days,1 month, and 3 months—in the coronary arteries of adult miniature swine.Forty (40) animals were studied at each time interval (10 animals perdose). Each animal received one drug-coated stent and one control stent.The control stent contained no drug. Table 8 shows the dosing scheme forswine efficacy study. TABLE 8 Dose group Dose group Dose group Dosegroup 1 (μg) 2 (μg) 3 (μg) 4 (μg) A-179578 per 15 45 150 400 stentA-179578 per  1  3  10  27 mm of stent

[0130] Potential local tissue toxicity was assessed at all timeintervals by examining histopathologic changes in the stented region,adjacent coronary segments, perivascular tissue, and subservedmyocardium. The mortality, angiographic implant and restudy data,histomorphometry data, and stent site histopathology were studied

[0131] Three-Day Group

[0132] Histopathology in combination with scanning electron microscopyprovided information regarding the short-term response to the implantedstent. The responses were similar in the control group and all dosegroups, and the responses involved compression of the tunica mediawithout remarkable necrosis, an accumulation of thrombus andinflammatory cells mostly localized to the stent struts, and earlyevidence of endothelial recovery and smooth muscle cell invasion of thethin mural thrombi. There were no extensive thrombi or remarkableintramural hemorrhages. The adventitia in some samples displayed eitherfocal or diffuse inflammatory infiltrates, and occasionally, there wasplugging or congestion of the vasa vasora. There was no evidence ofmedial necrosis in any sample.

[0133] Scanning electron microscopy showed similar appearance of theluminal surface three days after the implant of the coronary stent inall dose groups. The shape of the stent was clearly embedded in a thinlayer of tissue. The endothelium was intact between the struts and evenover the struts; a confluent or nearly confluent layer ofendothelial-like cells had covered the luminal surface. There werescattered adherent platelets, platelet microthrombi, and leukocytes overthe stents and on the intact remnant endothelium in the inter-strutspaces. In arteries with more severe stent-induced vessel damage, therewere more substantial mural thrombi, but the extent of endothelialrecovery over the stent struts did not appear retarded, regardless ofthe dosage of A-179578.

[0134] One-Month Group

[0135] The histomorphometry data for the one-month series indicated asignificant inhibitory effect of locally eluted A-179578 on neointimaformation in stented coronary arteries of swine. Intima area normalizedto injury score was significantly decreased for dose groups 3 and 4 (10and 27 μg/mm) as compared with the control; there were also trends fordecreases in absolute intima area and intima thickness for both dosegroups 3 and 4 as compared with the control, and a tendency towardsdecreased histologic % stenosis for dose group 3 as compared with thecontrol.

[0136] The control stents displayed morphology typical of stentsimplanted in coronary arteries of Yucatan miniature swine at one month.The tunica media was compressed or thinned without necrosis subjacent toprofiles of stent struts; there were only occasional inflammatoryinfiltrates; and the neointima ranged in size from relatively thin tomoderately thin, and were composed of spindle-shaped and stellate cellsin an abundant extracellular matrix, with only rare small foci offibrinoid material around the profiles of the stent struts. Thedrug-coated stents showed similar compression of the tunica mediawithout any substantial necrosis at any dose; like control devices,there was little inflammation present. The neointima was notably thinnerin dose groups 3 and 4, in some cases being composed of only a fewlayers of cells. In all dose groups, there were substantial numbers ofsamples in which moderately sized fibrinoid deposits and inspisatedthrombi were observed in the deep neointima. These were usuallyassociated with the stent struts but sometimes extended between strutprofiles. However, in no case was there exposure of thrombus on theluminal surface, as the deposits were encapsulated within fibrocellulartissue and covered with a flattened layer of periluminalendothelial-like cells.

[0137] Scanning electron microscopy confirmed that a confluent layer ofendothelial or endothelial-like cells covered the entire stentedsurface, and there was no difference between drug-coated stents andcontrol stents in terms of adherence of blood elements; leukocytes werepresent in approximately equal numbers in all groups. These findingsdemonstrate that while A-179578 was associated with decreased neointimaformation and persistent mural thrombi, sufficient vessel wall healingin response to stent injury had occurred within one month after thestent had been implanted. This vessel wall healing had rendered theluminal surface non-reactive for platelet adhesion and thrombusformation, and minimally reactive for leukocyte adherence. Additionally,there was no evidence of vessel wall toxicity even at the highest dose(27 μg/mm), as there was no medial necrosis or stent malapposition.

[0138] Three-Month Group

[0139] There were no significant differences between the dose groups forany histomorphometric parameters of stented coronary arterial dimensionin the three-month period of the study. However, there were weak trendsfor decreases in the two primary variables describing neointimaformation—the cross-sectional area and the % area stenosis of the lumen.

[0140] The histopathologic appearance of the control stents in the swinecoronary artery samples at three months after the implant appearedsimilar to that of the controls from the one-month group, and similar tothose of all the groups in the three-month period. All samples showedfibrocellular neointima formation with mostly spindle-shaped smoothmuscle-like cells in the neointima and a confluent squamous periluminalcell layer. There were no intramural hemorrhages or persistent fibrinoiddeposits in the neointima; however some samples, particularly those withthicker neointima, showed evidence of prior thrombus accumulation andsubsequent organization in the form of neovascularization in theneointima. On occasion, samples showed evidence of moderate to severeinflammatory reactions localized to the stent struts, associated withdestruction of the tunica media architecture. These were most oftenassociated with thicker neointima as well. However, these were few innumber and were found in the control group as well as in the drug-coatedstent groups. It is presumed that these represented eitheranimal-specific generalized reactions to the implanted stent, evidenceof contamination of the stent, or some combination of these two factors,and is commonly found at an incidence of about 10-15% in the studies ofstent implants in swine coronary arteries. There was no evidence ofnecrosis of the tunica media or separation of the media from the stentin any sample. The adventitia of most three-month implants appeared tohave somewhat greater neovascularization than did the one-monthimplants, but this did not appear related to control or test stentgroup. Scanning electron microscopy demonstrated confluent endotheliumwith rare adherent blood cells in the control group and all dose groups.

CONCLUSIONS

[0141] The stent coated with A-179578 reduced in-stent neointimaformation in swine coronary arteries and provided clear evidence of abiologic drug effect (unresorbed thrombus/fibrin deposits of neointima)at one month. There was a weak tendency for the stent coated withA-179578 to show a persistent inhibitory effect at the longer-term timeinterval of three months. There was no local coronary arterial walltoxicity in the form of medial necrosis or stent malappositionassociated with any dose group, including the highest dose ofapproximately 27 μg/mm stent length at any time interval examined. Allstents were well incorporated into the tissue, and there was evidence ofstable healing responses in the form of fibrocellular neointimalincorporation and endothelial coverage at the one-month interval and atthe three-month interval. The trend towards a sustained inhibitoryeffect at three months after the stent was implanted in this animal issurprising and provides evidence for potentially persistent effects inpreventing clinical restenosis resulting from implanted stents.

[0142] It is understood that the foregoing detailed description andaccompanying examples are merely illustrative and are not to be taken aslimitations upon the scope of the invention, which is defined solely bythe appended claims and their equivalents. Various changes andmodifications to the disclosed embodiments will be apparent to thoseskilled in the art. Such changes and modifications, including withoutlimitation those relating to the chemical structures, substituents,derivatives, intermediates, syntheses, formulations and/or methods ofuse of the invention, may be made without departing from the spirit andscope thereof.

What is claimed is:
 1. A medical device comprising a supportingstructure and the therapeutic substance

or a pharmaceutically acceptable salt or prodrug thereof and at leastone other therapeutic substance selected from the group consisting ofanti-proliferative agents, anti-platelet agents, anti-inflammatoryagents, anti-thrombotic agents, thrombolytic agents, cytotoxic drugs,agents that inhibit cytokine or chemokine binding, cellde-differentiation inhibitors, anti-lipaedemic agents, matrixmetalloproteinase inhibitors, and cytostatic drugs.
 2. The medicaldevice of claim 1, wherein said anti-proliferative agent is ananti-mitotic agent.
 3. The medical device of claim 2, wherein saidanti-mitotic agent is selected from the group consisting of vincaalkaloids, anti-mitotic alkylating agents, and anti-mitotic metabolites.4. The medical device of claim 1, wherein said anti-platelet agent isselected from the group consisting of agents that inhibit adhesion ofplatelets, agents that inhibit aggregation of platelets, and agents thatinhibit activation of platelets.
 5. The medical device of claim 1,wherein said anti-inflammatory agent is estradiol.
 6. The medical deviceof claim 1, wherein said anti-inflammatory agent is dexamethasone. 7.The medical device of claim 1, wherein said supporting structure isselected from the group consisting of coronary stents, peripheralstents, catheters, arterio-venous grafts, by-pass grafts, and drugdelivery balloons used in the vasculature.
 8. The medical device ofclaim 1, wherein said supporting structure further comprises a coating,said coating containing said therapeutic substances.
 9. The medicaldevice of claim 8, wherein said coating is polymeric.
 10. The medicaldevice of claim 1, wherein said therapeutic substance is

or a pharmaceutically acceptable salt or prodrug thereof.
 11. Themedical device of claim 1, wherein said therapeutic substance is

or a pharmaceutically acceptable salt or prodrug thereof.
 12. A medicaldevice comprising a supporting structure and the therapeutic substance

or a pharmaceutically acceptable salt or prodrug thereof and at leastone other therapeutic substance.
 13. The medical device of claim 12,wherein said at least one other therapeutic substance is a drug selectedfrom the group consisting of anti-proliferative agents, anti-plateletagents, anti-inflammatory agents, anti-thrombotic agents, thrombolyticagents, cytotoxic drugs, agents that inhibit cytokine or chemokinebinding, cell de-differentiation inhibitors, anti-lipaedemic agents,matrix metalloproteinase inhibitors, and cytostatic drugs.
 14. Themedical device of claim 13, wherein said anti-inflammatory agent isselected from the group consisting of estradiol and dexamethasone. 15.The medical device of claim 12, wherein said anti-lipaedemic agent isfenofibrate and said matrix metalloproteinase inhibitor is batimistat.16. The medical device of claim 12, wherein said supporting structurefurther comprises a coating, said coating containing said therapeuticsubstances.
 17. The medical device of claim 12, wherein said supportingstructure is selected from the group consisting of coronary stents,peripheral stents, catheters, arterio-venous grafts, by-pass grafts, anddrug delivery balloons used in the vasculature.
 18. A medical devicecomprising a supporting structure having a coating on the surfacethereof, said coating containing the therapeutic substance

or a pharmaceutically acceptable salt or prodrug thereof and at leastone drug selected from the group consisting of anti-proliferativeagents, anti-platelet agents, anti-inflammatory agents, anti-thromboticagents, thrombolytic agents, cytotoxic drugs, agents that inhibitcytokine or chemokine binding, cell de-differentiation inhibitors,anti-lipaedemic agents, matrix metalloproteinase inhibitors, andcytostatic drugs.
 19. The medical device of claim 18, wherein saidanti-proliferative agent is an anti-mitotic agent.
 20. The medicaldevice of claim 19, wherein said anti-mitotic agent is selected from thegroup consisting of vinca alkaloids, anti-mitotic alkylating agents, andanti-mitotic metabolites.
 21. The medical device of claim 18, whereinsaid anti-platelet agent is selected from the group consisting of agentsthat inhibit adhesion of platelets, agents that inhibit aggregation ofplatelets, and agents that inhibit activation of platelets.
 22. Themedical device of claim 18, wherein said anti-inflammatory agent isestradiol.
 23. The medical device of claim 18, wherein saidanti-inflammatory agent is dexamethasone.
 24. The medical device ofclaim 18, wherein said supporting structure is selected from the groupconsisting of coronary stents, peripheral stents, catheters,arterio-venous grafts, by-pass grafts, and drug delivery balloons usedin the vasculature.
 25. The medical device of claim 18, wherein saidcoating is polymeric.
 26. The medical device of claim 25, wherein saidpolymeric coating is biostable.
 27. The medical device of claim 25,wherein said polymeric coating is biodegradable.
 28. The medical deviceof claim 18, wherein said therapeutic substance is

or a pharmaceutically acceptable salt or prodrug thereof.
 29. Themedical device of claim 18, wherein said therapeutic substance is

or a pharmaceutically acceptable salt or prodrug thereof.
 30. A medicaldevice comprising a supporting structure having a coating on the surfacethereof, said coating containing the therapeutic substance

or a pharmaceutically acceptable salt or prodrug thereof and at leastone drug selected from the group consisting of anti-proliferativeagents, anti-platelet agents, anti-inflammatory agents, anti-thromboticagents, thrombolytic agents, cytotoxic drugs, agents that inhibitcytokine or chemokine binding, cell de-differentiation inhibitors,anti-lipaedemic agents, matrix metalloproteinase inhibitors, andcytostatic drugs.
 31. The medical device of claim 30, wherein saidanti-proliferative agent is an anti-mitotic agent.
 32. The medicaldevice of claim 31, wherein said anti-mitotic agent is selected from thegroup consisting of vinca alkaloids, anti-mitotic alkylating agents, andanti-mitotic metabolites.
 33. The medical device of claim 30, whereinsaid anti-platelet agent is selected from the group consisting of agentsthat inhibit adhesion of platelets, agents that inhibit aggregation ofplatelets, and agents that inhibit activation of platelets.
 34. Themedical device of claim 30, wherein said anti-inflammatory agent isestradiol.
 35. The medical device of claim 30, wherein saidanti-inflammatory agent is dexamethasone.
 36. The medical device ofclaim 30, wherein said supporting structure is selected from the groupconsisting of coronary stents, peripheral stents, catheters,arterio-venous grafts, by-pass grafts, and drug delivery balloons usedin the vasculature.
 37. The medical device of claim 30, wherein saidcoating is polymeric.
 38. The medical device of claim 37, wherein saidpolymeric coating is biostable.
 39. The medical device of claim 37,wherein said polymeric coating is biodegradable.
 40. A medical devicecomprising a supporting structure capable of containing or supporting apharmaceutically acceptable carrier or excipient, said carrier orexcipient containing the therapeutic substance

or a pharmaceutically acceptable salt or prodrug thereof and at leastone other therapeutic substance selected from the group consisting ofanti-proliferative agents, anti-platelet agents, anti-inflammatoryagents, anti-thrombotic agents, thrombolytic agents, cytotoxic drugs,agents that inhibit cytokine or chemokine binding, cellde-differentiation inhibitors, anti-lipaedemic agents, matrixmetalloproteinase inhibitors, and cytostatic drugs.
 41. The medicaldevice of claim 40, wherein said anti-proliferative agent is ananti-mitotic agent.
 42. The medical device of claim 41, wherein saidanti-mitotic agent is selected from the group consisting of vincaalkaloids, anti-mitotic alkylating agents, and anti-mitotic metabolites.43. The medical device of claim 40, wherein said anti-platelet agent isselected from the group consisting of agents that inhibit adhesion ofplatelets, agents that inhibit aggregation of platelets, and agents thatinhibit activation of platelets.
 44. The medical device of claim 40,wherein said anti-inflammatory agent is estradiol.
 45. The medicaldevice of claim 40, wherein said anti-inflammatory agent isdexamethasone.
 46. The medical device of claim 40, wherein saidsupporting structure includes a framework in the form of a stent. 47.The medical device of claim 40, wherein said supporting structure isselected from the group consisting of coronary stents, peripheralstents, catheters, arterio-venous grafts, by-pass grafts, and drugdelivery balloons used in the vasculature.
 48. The medical device ofclaim 40, wherein said supporting structure further comprises a coating,said coating containing said therapeutic substances.
 49. The medicaldevice of claim 48, wherein said coating is polymeric.
 50. The medicaldevice of claim 49, wherein said polymeric coating is biostable.
 51. Themedical device of claim 49, wherein said polymeric coating isbiodegradable.
 52. The medical device of claim 40, wherein saidsupporting structure includes a polymeric framework containing saidtherapeutic substances.
 53. The medical device of claim 40, wherein saidpolymeric framework is biodegradable.
 54. The medical device of claim40, wherein said anti-lipaedemic agent is fenofibrate.
 55. The medicaldevice of claim 40, wherein said matrix metalloproteinase inhibitor isbatimistat.